Modulation of cultured mouse leydig cells adenylate cyclase by forskolin and hCG

Lefèvre, Annick; Finaz, Catherine; Berthelon, M.; Saez, J.M.

doi:10.1016/0303-7207(85)90164-9

Cited by 16 publications

(4 citation statements)

References 26 publications

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“…Another AC activator, forskolin, mediated its stimulating effect on steroidogenesis through activation of an AC catalytic subunit [7]. This preparation also caused an increase of testosterone production by murine Leydig's cells (F7.26=8.3, p<0.001), but its effect on steroidogenesis was less pronounced than that of chorionic gonadotropin and cholera toxin.…”

Section: Resultsmentioning

confidence: 95%

Genetic basis of adenylatecyclase regulation of testosterone production of Leydig's cells in laboratory mice

Осадчук

Svechnikov

1994

Bull Exp Biol Med

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The function of Leydig's cells during stimulation of various components of the adenylatecyclase system was studied in inbred mice of various lines. Mice of different lines were found to differ markedly in their production of testosterone during stimulation of a crude Leydig's cell suspension with ascending concentrations of chorionic gonadotropin, cholera toxin, forskolin, and dibutyryl-cAMP. The ranking of the lines according to the maximal production of testosterone was virtually the same during exposure to steroidogenesis activators.

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Section: Resultsmentioning

confidence: 95%

Genetic basis of adenylatecyclase regulation of testosterone production of Leydig's cells in laboratory mice

Осадчук

Svechnikov

1994

Bull Exp Biol Med

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“…Prior to biochemical testing, the cells were incubated 7 days in a serum-free chemically defined medium (6). Binding of highly purified hCG (CR 123, 12,750 international units/mg, prepared by R. E. Canfield) was studied as described (6) after iodination of the hormone by the lactoperoxidase method (14) to a specific activity of 50-60 ,tCi/sg (1 Ci = 37 GBq).…”

Section: Methodsmentioning

confidence: 99%

“…Fresh Leydig cells, isolated from the testes of 6-week-old BALB/c mice, were purified and maintained in primary culture as previously described (6,12). Ouabain at 20,4M will kill fresh Leydig cells but not the LK clone.…”

Section: Methodsmentioning

confidence: 99%

“…Leydig cells in primary culture have been used to study the mechanism of action of gonadotropin, but testosterone production is not sustained more than a few days (2)(3)(4)(5)(6). Clonal cell lines derived from mouse (7)(8)(9) and rat (10) Leydig cell tumors, although they may retain LH/hCG receptors, are unable to pursue steroidogenesis beyond progesterone synthesis.…”

Section: For Review)mentioning

confidence: 99%

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Construction of a Leydig cell line synthesizing testosterone under gonadotropin stimulation: a complex endocrine function immortalized by cell hybridization.

Finaz¹,

Lefèvre²,

Dampfhoffer³

1987

Proc. Natl. Acad. Sci. U.S.A.

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Hybridization between a mouse Leydig tumor cell line, MA-l0, which produces cyclic AMP and progesterone under human chorionic gonadotropin (hCG) stimulation, and freshly isolated mouse Leydig cells gave rise to 54 hybrid clones, one of which, LK17, was capable of hCGstimulated testosterone production. Subcloning of this hybrid resulted in the emergence of a subclone, K9, whose testosterone production is more than 10 times that of parent clone LK17, after hCG stimulation, with an ED50 of 37 pM. Testosterone synthesis by K9 cells was multiplied by 25 after gonadotropin stimulation, and binding of hCG declined after prolonged exposure to the hormone. These similarities with murine Leydig cells in primary culture make the K9 clone an attractive alternative for physiological studies.Production of testosterone by Leydig cells is the outcome of a complex steroidogenic pathway, initiated by the cleavage of the side chain of cholesterol. This step is triggered by the binding of luteinizing gonadotropin to luteinizing hormone! human chorionic gonadotropin (LH/hCG) receptors located on the cell plasma membrane, followed by the activation of adenylate cyclase and protein kinase (see ref. 1 for review). Leydig cells in primary culture have been used to study the mechanism of action of gonadotropin, but testosterone production is not sustained more than a few days (2-6). Clonal cell lines derived from mouse (7-9) and rat (10) Leydig cell tumors, although they may retain LH/hCG receptors, are unable to pursue steroidogenesis beyond progesterone synthesis. Using somatic cell hybridization between the MA-10 mouse Leydig tumor cell line obtained by Ascoli (11) and freshly isolated mouse Leydig cells, we have immortalized the full steroidogenic function of the latter. The strategy used for obtaining this testosterone-producing gonadotropin-sensitive permanent cell line is described below. MATERIALS AND METHODSParental Cells. A clonal mouse Leydig tumor cell line, , was kindly provided by Mario Ascoli (Nashville, TN). This cell line responds to hCG stimulation by an increased secretion of progesterone and 20a-dehydroprogesterone. It was made hypoxanthine/guanine phosphoribosyltransferase negative (HGPRT-) by mutation using ethyl methanesulfonate, added at 300 ,4g/ml for 14 hr to cells grown to confluence in Dulbecce's modified Eagle's medium containing 15% fetal calf serum. The treated cells were allowed to recover 7 days in the initial medium, and HGPRT-cells were then selected by addition of 100 ,uM 6-thioguanine. Mutant colonies appeared 3 weeks later. A mutant clone, LK, was chosen for its endocrinological similarity to the MA-10 parent cell line-namely, the ability to respond to hCG stimulation by an increased progesterone production.Fresh Leydig cells, isolated from the testes of 6-week-old BALB/c mice, were purified and maintained in primary culture as previously described (6, 12). Ouabain at 20,4M will kill fresh Leydig cells but not the LK clone.Cell Hybridization and Subcloning. Twelve hours after plating of the ...

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Systematic shut-off of the hormone receptors in intraspecific adrenal X Leydig cell hybrids

Lefèvre

Finaz

1986

Experimental Cell Research

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Modulation of cultured mouse leydig cells adenylate cyclase by forskolin and hCG

Cited by 16 publications

References 26 publications

Genetic basis of adenylatecyclase regulation of testosterone production of Leydig's cells in laboratory mice

Genetic basis of adenylatecyclase regulation of testosterone production of Leydig's cells in laboratory mice

Construction of a Leydig cell line synthesizing testosterone under gonadotropin stimulation: a complex endocrine function immortalized by cell hybridization.

Systematic shut-off of the hormone receptors in intraspecific adrenal X Leydig cell hybrids

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